Vishal Gaurav - Isolation and Propagation of High Secondary Metabolite-Accumulating Plant Cell Cultures

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      Publication Details (including relevant citation   information):

      Authors: Vishal Gaurav, Susan C. Roberts

      Publication date: 2009

      Conference name: American Institute of Chemical Engineers (AIChE)   Proceedings

      Publisher: 2009 Annual Meeting, Nashville, TN


      Plant cell culture technology is used for the production of many   important natural products including pharmaceuticals, flavors,   colors and other specialty chemicals. While more than 25% of all   the pharmaceutical prescribed in the US are plant derived, less   than 2% of all the plant species have been investigated for   potential medical applications. A prominent example of plant cell   culture technology, in both research and industrial applications,   is Taxus spp. for production of anti-cancer agent paclitaxel   (Taxol®). There are, however, limitations associated with this   technology that prevent widespread industrial use. Cell-cell   heterogeneity is a typical feature of these cultures, which   results in unpredictable shifts in metabolite accumulation over   time and within cultures, thus leading to low and variable yields   of these products. We have developed a novel flow cytometric   method for immunostaining and isolating high paclitaxel   -accumulating sub-populations from Taxus cell cultures.   Florescence-activated cell sorting (FACS) was used to aseptically   sort these cultures into high- and low- paclitaxel producing   sub-populations. Methods to propagate single cell suspension   cultures from these isolated populations were subsequently   developed. Sorted suspension cultures were scaled-up, and their   growth, viability, size distribution and paclitaxel accumulation   were analyzed over time. Aggregate and cell size distribution was   determined using Coulter Counter technology and quantification of   paclitaxel accumulation in single cell cultures was performed   using flow cytometry. Culture conditions (e.g., seeding density,   elicitation, media composition, etc.) were optimized for cell   growth and production. This technology, through isolation of   high-accumulators and establishment of elite cell lines, will   provide a more efficient and economically viable method for   production of paclitaxel and other plantderived natural products   from suspension culture.

      Address (URL): dary_Metabolite-Accumulating_Plant_Cell_Cultures.pdf